bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 2, 2025
ABSTRACT
Rhodamine
dyes
are
widely
used
fluorophores
in
super-resolution
fluorescence
imaging
due
to
their
exceptional
optical
properties
and
“aggregation-disaggregation”
induced
fluorogenic
activation.
However,
excessive
lipophilicity
often
reduces
brightness
aqueous
environments
causes
off-target
staining,
limiting
effectiveness
high-resolution
imaging.
To
address
these
challenges,
we
introduce
an
ether-decorated
N-terminal
modification
strategy
for
rhodamine
silicon-rhodamine
(Si-rhodamine),
replacing
conventional
N-alkyl
groups.
The
ether
chains
enhance
water
solubility,
decrease
aggregate
size,
improve
fluorogenicity
across
a
wide
concentration
range.
Their
flexible,
hydrophilic
structure
forms
protective
shield
around
the
xanthene
core,
minimizing
dye-water
interactions
reducing
quenching.
Additionally,
inductive
effect
of
decreases
electron-donating
strength
amino
groups,
suppressing
quenching
caused
by
twisted
intramolecular
charge
transfer
(TICT).
These
modifications
collectively
increase
quantum
yields
ER
ESiR
from
0.35
0.19
(for
tetraethyl-substituted
analogs)
0.70
0.41,
respectively.
Probes
derived
exhibit
outstanding
fluorogenicity,
enhanced
signal-to-noise
ratios,
improved
resolution
complex
environments,
demonstrating
superior
performance
advanced
techniques
such
as
structured
illumination
microscopy
(SIM),
stimulated
emission
depletion
(STED)
microscopy,
single-molecule
localization
(SMLM).
This
work
introduces
innovative
fluorophore
design,
offering
significant
advancements
applications.
Chemistry of Materials,
Год журнала:
2023,
Номер
36(2), С. 949 - 958
Опубликована: Дек. 29, 2023
The
confocal
and
super-resolution
imaging
has
become
routine
for
the
visible
spectral
region,
but
not
deep
near-infrared
(NIR)
region
(800–1000
nm)
due
to
lack
of
bright
ultraphotostable
fluorophores.
We
devised
a
premium
fluorochromic
scaffold
(EC5)
spectrally
active
in
this
further
optimized
it
by
rational
systematic
molecular
engineering
over
push–pull
headgroups.
EC5j
maximally
absorbs/emits
at
835/873
nm
offers
superior
brightness
38070
cm–1
M–1
photostability.
Using
two
home-built
microscopes
with
deep-NIR
capability,
i.e.,
structured-illumination
microscopes,
we
demonstrated
that
2D
3D-microscopy
is
now
practical
EC5j.
In
particular,
EC5j-adsorbed
polystyrene
beads
furnished
notable
spatial
resolution
175
structure-illumination
microscopy
(SIM)
imaging,
as
compared
theoretical
diffraction
limit
336
nm.
Proof-of-concept
applications
included
live-cell
mitophagy,
SIM
mitochondria,
deconvolution-based
sectioning
cell
structures
brain
vasculature.
revolutionary
expansion
window
brings
field
new
tools
insights.
Organic
fluorophores
with
near-infrared
(NIR)
emissions
and
reduced
molecular
weights
are
crucial
for
advancing
bioimaging
biosensing
technologies.
Traditional
methods,
such
as
conjugation
expansion
heteroatom
engineering,
often
fail
to
reduce
fluorophore
size
without
sacrificing
NIR
emission
properties.
Addressing
this
challenge,
our
study
utilized
computational
screening
structure-property
relationship
analysis
establish
comprehensive
design
principles
compact,
single-benzene-based
fluorophores.
These
newly
developed
not
only
exhibit
above
700
nm
but
also
maintain
under
200
g/mol,
approximately
25%
of
that
Cy7.
Additionally,
they
display
unique
environmental
sensitivity—non-emissive
in
aqueous
solutions
highly
emissive
lipid
environments.
This
property
significantly
enhances
their
utility
live
cell
imaging
by
enabling
wash-free
applications.
Our
findings
mark
a
substantial
breakthrough
paving
the
way
more
efficient
adaptable
methodologies.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2025,
Номер
unknown
Опубликована: Янв. 2, 2025
ABSTRACT
Rhodamine
dyes
are
widely
used
fluorophores
in
super-resolution
fluorescence
imaging
due
to
their
exceptional
optical
properties
and
“aggregation-disaggregation”
induced
fluorogenic
activation.
However,
excessive
lipophilicity
often
reduces
brightness
aqueous
environments
causes
off-target
staining,
limiting
effectiveness
high-resolution
imaging.
To
address
these
challenges,
we
introduce
an
ether-decorated
N-terminal
modification
strategy
for
rhodamine
silicon-rhodamine
(Si-rhodamine),
replacing
conventional
N-alkyl
groups.
The
ether
chains
enhance
water
solubility,
decrease
aggregate
size,
improve
fluorogenicity
across
a
wide
concentration
range.
Their
flexible,
hydrophilic
structure
forms
protective
shield
around
the
xanthene
core,
minimizing
dye-water
interactions
reducing
quenching.
Additionally,
inductive
effect
of
decreases
electron-donating
strength
amino
groups,
suppressing
quenching
caused
by
twisted
intramolecular
charge
transfer
(TICT).
These
modifications
collectively
increase
quantum
yields
ER
ESiR
from
0.35
0.19
(for
tetraethyl-substituted
analogs)
0.70
0.41,
respectively.
Probes
derived
exhibit
outstanding
fluorogenicity,
enhanced
signal-to-noise
ratios,
improved
resolution
complex
environments,
demonstrating
superior
performance
advanced
techniques
such
as
structured
illumination
microscopy
(SIM),
stimulated
emission
depletion
(STED)
microscopy,
single-molecule
localization
(SMLM).
This
work
introduces
innovative
fluorophore
design,
offering
significant
advancements
applications.
